The Comparative Study of the Mouse Osteoblast Response to Two Different Platelet-rich Fibrins with Low Speed Centrifugation Concept

Domprapagon Patthapong, Tachasuttirut Kathawut, Permpanich Piyanuj

Published Online: Apr 18, 2024 DOI 10.12982/OSR.2024.022

Objectives: To compare the response of mouse osteoblast (MC3T3-E1 cells) to advanced platelet-rich fibrin (APRF) and advanced platelet-rich fibrin plus (APRF+).

Methods: Blood was collected from eight volunteers, 25 - 38 years of age (four males and four females) to prepare platelet concentrates APRF (1,300 rpm, 14 minutes), APRF+ (1,300 rpm, 8 minutes). The exudates were collected from both platelet concentrates at day 1, 3, 7 and 14. The level of TGF-β1 from exudates were quantified using an ELISA. MC3T3-E1 cells were cultured with the exudates. The cultured cells were tested with MTT assays, alkaline phosphatase (ALP) staining and mineralization, which were analyzed on day 7 and again on day 14.

Results: APRF and APRF+ continuously released TGF-β1 during 14-days period. Only exudates collected at day 1 showed significantly difference of ALP staining between APRF and APRF+ group on day 14 of observation. On day 7 of mineralization assays, cells treated with exudates from APRF+ collected at day 14 resulted in the highest level of mineralization within APRF+ group. On day 14 of mineralization assays, cells treated with exudates from APRF collected at day 7 showed the highest level of mineralization within APRF group.

Conclusions: APRF+ released TGF-β1 at day 14 significantly higher than day 1. On day 14 of mineralization, cells treated with exudates from APRF+ collected at day 14 showed significantly higher mineralization than APRF collected at the same time point.

advanced platelet-rich fibrin advanced platelet-rich fibrin plus low speed centrifugation concept mineralization platelet concentrates
  1. Kinane DF, Stathopoulou PG, Papapanou PN. Periodontal diseases. Nat Rev Dis Primers. 2017;3:17038.
  2. Chen F, Zhang J, Zhang M, An Y, Chen F, Wu ZF. A review on endogenous regenerative technology in periodontal regenerative medicine. Biomaterials. 2010;31(31):7892-927.
  3. Ezzatt OM. Autologous Platelet Concentrate Preparations in Dentistry. Biomed J Sci Tech Res. 2018;8:6712-20.
  4. Marx RE, Carlson ER, Eichstaedt RM, Schimmele SR, Strauss JE, Georgeff KR. Platelet-rich plasma: growth factor enhancement for bone grafts. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 1998;85(6):638-46.
  5. Landesberg R, Moses M, Karpatkin M. Risks of using platelet rich plasma gel. J Oral Maxillofac Surg. 1998;56(9):1116-7.
  6. Dohan DM, Choukroun J, Diss A, Dohan SL, Dohan AJ, Mouhyi J, et al. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part I: technological concepts and evolution. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2006;101(3):e37-e44.
  7. Dohan DM, de Peppo GM, Doglioli P, Sammartino G. Slow release of growth factors and thrombospondin-1 in Choukroun's platelet-rich fibrin (PRF): a gold standard to achieve for all surgical platelet concentrates technologies. Growth Factors. 2009;27(1):63-9.
  8. Choukroun J, Diss A, Simonpieri A, Girard MO, Schoeffler C, Dohan SL, et al. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part V: histologic evaluations of PRF effects on bone allograft maturation in sinus lift. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2006;101(3):299-303.
  9. Agarwal A, Gupta ND, Jain A. Platelet rich fibrin combined with decalcified freeze-dried bone allograft for the treatment of human intrabony periodontal defects: a randomized split mouth clinical trail. Acta Odontol Scand. 2016;74(1):36-43.
  10. Jankovic S, Aleksic Z, Klokkevold P, Lekovic V, Dimitrijevic B, Barrie Kenney EB, et al. Use of platelet-rich fibrin membrane following treatment of gingival recession: a randomized clinical trial. Int J Periodontics Restorative Dent. 2012;32(2):41-50.
  11. Ghanaati S, Booms P, Orlowska A, Kubesch A, Lorenz J, Rutkowski J, et al. Advanced platelet-rich fibrin: a new concept for cell-based tissue engineering by means of inflammatory cells. J Oral Implantol. 2014;40(6):679-89.
  12. Bagdadi KEI, Kubesch A, Yu X, Maawi SAI, Orlowska A, Dias A, et al. Reduction of relative centrifugal forces increases growth factor release within solid platelet-rich-fibrin (PRF)-based matrices: a proof of concept of LSCC (low speed centrifugation concept). Eur J Trauma Emerg Surg. 2019;45(3):467-79.
  13. Fujioka-Kobayashi M, Miron RJ, Hernandez M, Kandalam U, Zhang Y, Choukroun J. Optimized platelet-rich fibrin with the low-speed concept: growth factor release, biocompatibility, and cellular response. J Periodontol. 2017;88(1):112-21.
  14. Fujioka-Kobayashi M, Miron RJ. Biological Components of Platelet Rich Fibrin: Growth Factor Release and Cellular Activity.In: Miron RJ, Choukroun J, editors. Platelet Rich Fibrin in Regenerative Dentistry: Biological Background and Clinical Indications. 1st ed. Wiley-Blackwell; 2017.p.15-31.
  15. Choukroun J, Ghanaati S. Reduction of relative centrifugation force within injectable platelet-rich-fibrin (PRF) concentrates advances patients’own inflammatory cells, platelets and growth factors: the first introduction to the low speed centrifugation concept. Eur J Trauma Emerg Surg. 2017;44(1):87–95.
  16. Dohan DM, Del Corso M, Diss A, Mouhyi J, Charrier JB. Three-dimensional architecture and cell composition of a Choukroun's platelet-rich fibrin clot and membrane. J Periodontol. 2010;81(4):546-55.
  17. Sahni A, Francis CW. Vascular endothelial growth factor binds to fibrinogen and fibrin and stimulates endothelial cell proliferation. Blood. 2000;96(12):3772-8.
  18. Martino MM, Briquez PS, Ranga A, Lutolf MP, Hubbell JA. Heparin-binding domain of fibrin(ogen) binds growth factors and promotes tissue repair when incorporated within a synthetic matrix. Proc Natl Acad Sci U S A. 2013;110(12):4563-8.
  19. Kirchberg K, Lange TS, Klein EC, Jungtäubl H, Heinen G, Meyer-Ingold W, et al. Induction of β1 integrin synthesis by recombinant platelet-derived growth factor (PDGF-AB) correlates with an enhanced migratory response of human dermal fibroblasts to various extracellular matrix proteins. Exp Cell Res. 1995;220(1),29-35.
  20. Brem H, Kodra A, Golinko MS, Entero H, Stojadinovic O, Wang VM, et al. Mechanism of sustained release of vascular endothelial growth factor in accelerating experimental diabetic healing. J Invest Dermatol. 2009;129(9):2275-87.
  21. Kermani S, Megat Abdul Wahab R, Zarina Zainol Abidin I, Zainal Ariffin Z, Senafi S, Hisham Zainal Ariffin S. Differentiation capacity of mouse dental pulp stem cells into osteoblasts and osteoclasts. Cell J. 2014;16(1):31-42.
  22. Quarles LD, Yohay DA, Lever LW, Caton R, Wenstrup RJ. Distinct proliferative and differentiated stages of murine MC3T3-E1 cells in culture: an in vitro model of osteoblast development. J Bone Miner Res. 1992;7(6):683-92.
  23. He L, Lin Y, Hu X, Zhang Y, Wu H. A comparative study of platelet-rich fibrin (PRF) and platelet-rich plasma (PRP) on the effect of proliferation and differentiation of rat osteoblasts in vitro. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2009;108(5):707-13.
  24. Lundquist R, Dziegiel MH, Agren MS. Bioactivity and stability of endogenous fibrogenic factors in platelet-rich fibrin. Wound Repair Regen. 2008;16(3):356–63.

Domprapagon P, Tachasuttirut K, Permpanich P. The Comparative Study of the Mouse Osteoblast Response to Two Different Platelet-rich Fibrins with Low Speed Centrifugation Concept: Original articles. CM Dent J [Internet]. 2024 Apr 18 [cited 2024 Nov 18];45(1):39-46. Available from: https://www.dent.cmu.ac.th/cmdj/frontend/web/?r=site/viewarticle&id=213

Domprapagon, P., Tachasuttirut, K. & Permpanich, P. (2024). The Comparative Study of the Mouse Osteoblast Response to Two Different Platelet-rich Fibrins with Low Speed Centrifugation Concept. CM Dent J, 45(1), 39-46. Retrieved from: https://www.dent.cmu.ac.th/cmdj/frontend/web/?r=site/viewarticle&id=213

Domprapagon, P., Tachasuttirut Kathawut and Permpanich Piyanuj. 2024. "The Comparative Study of the Mouse Osteoblast Response to Two Different Platelet-rich Fibrins with Low Speed Centrifugation Concept." CM Dent J, 45(1), 39-46. https://www.dent.cmu.ac.th/cmdj/frontend/web/?r=site/viewarticle&id=213

Domprapagon, P. et al. 2024. 'The Comparative Study of the Mouse Osteoblast Response to Two Different Platelet-rich Fibrins with Low Speed Centrifugation Concept', CM Dent J, 45(1), 39-46. Retrieved from https://www.dent.cmu.ac.th/cmdj/frontend/web/?r=site/viewarticle&id=213

Domprapagon, P., Tachasuttirut, K. and Permpanich, P. "The Comparative Study of the Mouse Osteoblast Response to Two Different Platelet-rich Fibrins with Low Speed Centrifugation Concept", CM Dent J, vol.45, no. 1, pp. 39-46, Apr. 2024.

Domprapagon Patthapong, Tachasuttirut Kathawut, Permpanich Piyanuj "The Comparative Study of the Mouse Osteoblast Response to Two Different Platelet-rich Fibrins with Low Speed Centrifugation Concept." CM Dent J, vol.45, no. 1, Apr. 2024, pp. 39-46, https://www.dent.cmu.ac.th/cmdj/frontend/web/?r=site/viewarticle&id=213